title: Generation of a tightly regulated doxycycline-inducible model for studying mouse intestinal biology.
abstract: To develop a sensitive and inducible system to study intestinal biology, we generated a transgenic mouse model expressing the reverse tetracycline transactivator rtTA2-M2 under control of the 12.4 kb murine Villin promoter. The newly generated Villin-rtTA2-M2 mice were then bred with the previously developed tetO-HIST1H2BJ/GFP model to assess inducibility and tissue-specificity. Expression of the histone H2B-GFP fusion protein was observed exclusively upon doxycycline induction and was uniformly distributed throughout the intestinal epithelium. The Villin-rtTA2-M2 was also found to drive transgene expression in the developing mouse intestine. Furthermore, we could detect transgene expression in the proximal tubules of the kidney and in a population of alleged gastric progenitor cells. By administering different concentrations of doxycycline, we show that the Villin-rtTA2-M2 system drives transgene expression in a dosage-dependent fashion. Thus, we have generated a novel doxycycline-inducible mouse model, providing a valuable tool to study the effect of different gene dosages on intestinal physiology and pathology.
authors.affiliation: Department of Pathology, Josephine Nefkens Institute, Erasmus MC, Rotterdam, The Netherlands.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Roth
authors.forename: Patrick
authors.initials: P
authors.lastname: Franken
authors.forename: Wendy
authors.initials: W
authors.lastname: van Veelen
authors.forename: Lau
authors.initials: L
authors.lastname: Blonden
authors.forename: Lalini
authors.initials: L
authors.lastname: Raghoebir
authors.forename: Berna
authors.initials: B
authors.lastname: Beverloo
authors.forename: Ellen
authors.initials: E
authors.lastname: van Drunen
authors.forename: Ernst J
authors.initials: EJ
authors.lastname: Kuipers
authors.forename: Robbert
authors.initials: R
authors.lastname: Rottier
authors.forename: Riccardo
authors.initials: R
authors.lastname: Fodde
authors.forename: Ron
authors.initials: R
authors.lastname: Smits
mesh_terms: D000818:Animals; D004318:Doxycycline / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects*; D017930:Genes, Reporter / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000187:drug effects / Q000502:physiology*; D051379:Mice; D008822:Mice, Transgenic; D009928:Organ Specificity; D012106:Research; D017186:Titrimetry; D019076:Transgenes / Q000235:genetics
chemical_list: D004318:Doxycycline